CN1984922A

CN1984922A - Substituted prolines as inhibitors of hepatitis c virus NS3 serine protease

Info

Publication number: CN1984922A
Application number: CNA2005800240748A
Authority: CN
Inventors: A·阿拉萨潘; F·G·恩约洛吉; V·M·吉里亚瓦拉布汉
Original assignee: Schering Corp
Current assignee: Merck Sharp and Dohme Corp
Priority date: 2004-05-20
Filing date: 2005-05-18
Publication date: 2007-06-20
Also published as: EP1773868B1; MXPA06013404A; CA2566610A1; EP1773868A1; HK1099028A1; WO2005113581A1; ES2328596T3; DE602005015452D1; US7399749B2; JP2008502718A; US20050272663A1

Abstract

The present invention discloses compounds of formula (I) which have HCV protease inhibitory activity as well as methods for preparing such compounds. In another embodiment, the invention discloses pharmaceutical compositions comprising such compounds as well as methods of using them to treat disorders associated with the HCV protease.

Description

Substituted prolines as inhibitors of NS 3 serine protease of hepatitis C virus

Invention field

The present invention relates to new hepatitis C virus (" HCV ") proteinase inhibitor, contain the pharmaceutical composition of one or more described inhibitor, the method for the described inhibitor of preparation and the method for using described inhibitor for treating hepatitis C and relative disease.The invention also discloses and contain the compound of new proline(Pro) part in the P2 position as HCV NS3/NS4a serpin.The application requires the right of priority of the U.S. Provisional Patent Application 60/573,191 of application on May 20th, 2004.

Background of invention

Hepatitis C virus (HCV) is a kind of (+)-adopted single strand RNA virus is arranged, and it is non-A non-B hepatitis (NANBH), the especially main pathogens of blood dependency NANBH (BB-NANBH) (referring to international application published WO89/04669 and European patent application published EP381216).NANBH both had been different from the virus induction hepatopathy of other type, for example hepatitis A virus (HAV), hepatitis B virus (HBV), fourth type (δ) hepatitis virus (HDV), cytomegalovirus (CMV) and Epstein-Barr virus (EBV), the hepatopathy that also is different from other form, for example alcoholism hepatopathy and primary biliary cirrhosis.

Recently, identify, clone and expressed polypeptide processing and the necessary HCV proteolytic enzyme of virus replication (referring to for example United States Patent (USP) 5,712,145).This about 3000 amino acid whose polyproteins comprise nucleocapsid protein (C), envelope protein (E1 and E2) and several Nonstructural Protein (NS1, NS2, NS3, NS4a, NS5a and NS5b) from aminoterminal to carboxyl terminal.NS3 by genomic about 1893 nucleotide codings of HCV, and has 2 different structural domains: (a) the serine protease structural domain of being made up of about 200 N terminal amino acids for the albumen of about 68kda; (b) PROTEIN C end RNA dependency ATP enzymatic structure territory.Because the similarity of protein sequence, whole three-dimensional structure and catalyst mechanism, NS3 proteolytic enzyme is considered to a member of chymotrypsin protein enzyme family.The flexible proteolytic enzyme of other chymotrypsin protein sample enzyme, factor Xa, zymoplasm, trypsinase, plasmin, urokinase, tPA and PSA.HCV NS3 serine protease is relevant with the hydrolysis of NS5a/NS5b contact in NS3/NS4a, NS4a/NS4b, NS4b/NS5a with described polypeptide (polyprotein), therefore is responsible for generating four kinds of viral proteins during virus replication.This makes HCV NS3 serine protease become the antiviral chemotherapeutical target that attracts people's attention.The compounds of this invention can suppress this proteinoid enzyme.They can also regulate the processing of hepatitis C virus (HCV) polypeptide.

The NS4a albumen of having determined about 6kda polypeptide is the cofactor of NS3 serine protease.The NS3/NS4a serine protease carries out at intramolecularly (being cis) the automatic cutting of NS3/NS4a contact, and other cleavage site carries out in intermolecular (promptly trans).

Analysis to the natural cleavage site of HCV proteolytic enzyme discloses, and there is halfcystine in P1, and there is Serine in P1 ', and these residues are strict conservative in NS4a/NS4b, NS4b/NS5a and NS5a/NS5b contact.The NS3/NS4a contact comprises the Threonine of P1 and the Serine of P1 '.Cys on the NS3/NS4a → Thr replaces and is considered to the reason that this contact needs cis rather than trans processing.Referring to (1994) such as for example Pizzi Proc.Nat1.Acad. Sci (USA) 91: 888-892, Failla etc. (1996) Folding﹠amp; Design 1: 35-42.The NS3/NS4a cleavage site also more tolerates mutagenesis than other site.Referring to (1 994) such as for example Kollykhalov J.Virol. 68: 7525-7533.Find that also the acidic residues of cleavage site upstream is that effectively cutting is needed.Referring to (1994) such as for example Komoda J.Virol. 68: 7351-7357.

The HCV proteinase inhibitor of having reported comprise antioxidant (referring to international application published WO98/14181), some peptide class and peptide analogs (referring to international application published WO98/17679, Landro etc. (1997) Biochem. 36: 9340-9348, Ingallinella etc. (1998) Biochem. 37: 8906-8914, (1998) such as Llin à s-Brunet Bioorg.Med.Chem. Lett. 8: 1713-1718), based on the inhibitor (Martin etc. (1998) of 70 amino acid whose polypeptide eglin c Biochem. 37: 11459-11468), be selected from human pancreas's secretor type trypsin inhibitor (hPSTI-C3) and miniantibody storehouse (minibody repertoire) inhibitor affinity (Dimasi etc. (1997) (MBip) J.Virol., 71: 7461-7469), cV _HE2 (" camelized " variable region antibody fragment) (Martin etc. (1997) Protein Eng. 10: 607-614) with α l-chymotrypsin inhibitor (ACT) (Elzouki etc.) (1997) J.Hepat. 27: 42-28).Disclose design recently and be used for the ribozyme of selective destruction HCV RNA (referring to BioWorld Today 9 (217): 4 (on November 10th, 1998)).

In addition referring on April 30th, 1998 disclosed PCT announcement WO98/17679 (VertexPharmaceuticals Incorporated), disclosed WO98/22496 on May 28th, 1998 (F.Hoffmann-La Roche AG) and disclosed WO99/07734 on February 18th, 1999 (Boehringer Ingelheim Canada Ltd.).

HCV is relevant with liver cirrhosis, and brings out hepatocellular carcinoma.HCV the infected's prognosis is very poor at present.Because lack immunity or infect relevant exemption (remission) with HCV, the hepatitis that HCV infects compared with other form more is difficult to treatment.Present data shows that the survival rate in 4 years is lower than 50% behind the liver cirrhosis diagnosis.The five-year survival rate that is diagnosed as the patient who suffers from focal resectability hepatocellular carcinoma is 10-30%, and focal patient's that can not excision property hepatocellular carcinoma five-year survival rate is lower than 1%.

Referring to WO00/59929 (US6,608,027, transferee: Boehringer Ingelheim (Canada) Ltd.; Announced on October 12nd, 2000), it discloses the peptide derivant of following formula:

Referring to A.Marchetti etc., Synlett, S1, 1000-1002 (1999), it has introduced the synthetic method of the dicyclo analogue of HCV NS3 proteinase inhibitor.Compound with following formula structure is wherein disclosed:

Referring to W.Han etc., Bioorganic﹠amp; Medicinal Chem.Lett, (2000) 10, 711-713, it introduces the preparation method of some alpha-keto amide class that contains allyl group and ethyl functional group, α-ketone ester class and α-cyclohexadione compounds.

Referring to WO00/09558 (transferee: Boehringer Ingelheim Limited; Announced on February 24th, 2000), it discloses the peptide derivant of following formula:

Each variable defines in the document in the formula.Be an example of this series compound below:

Referring to WO00/09543 (transferee: Boehringer Ingelheim Limited; Announced on February 24th, 2000), it discloses the peptide derivant of following formula:

Referring to U.S.6,608,027 (Boehringer Ingelheim, Canada), it discloses the NS3 proteinase inhibitor of following type:

Each variable defines in the document in the formula.

The current therapy of hepatitis C comprises interferon-' alpha ' (INF _α) and the conjoint therapy that uses ribavirin and Interferon, rabbit.Referring to (1998) such as for example Beremguer Proc.Assoc.Am. Physicians 110 (2): 98-112.There is the low defective that continues response rate and side effect takes place often in these therapies.Referring to (1997) such as for example Hoofnagle N.Engl.J.Med. 336:347.At present, infection does not have vaccine to use for HCV.

Referring to the WO01/74768 that announces October 11 calendar year 2001 (transferee: VertexPharmaceuticals Inc), it discloses the compound (defining in the R such as the document) that has following general formula as the part of NS 3-serine protease inhibitors of hepatitis C virus:

The disclosed particular compound of above-mentioned WO01/74768 has following structural:

PCT announces the U.S. Patent application that waits for ratification 10/052 of WO01/77113, WO01/081325, WO02/08198, WO02/08256, WO02/08187, WO02/08244, WO02/48172, WO02/08251 and application on January 18th, 2002,386, various types of peptide and/or other compounds as hepatitis C virus NS-3 serpin are disclosed.These apply for that disclosed content is attached to herein by reference.

Infection needs new methods of treatment for HCV.Need can be used for treating, preventing or improve the compound of one or more symptoms of hepatitis C.

Need to treat, prevent or improve the method for one or more symptoms of hepatitis C.

Need to regulate serine protease, the especially method of HCVNS3/NS4a serine protease with compound provided herein.

Need to regulate HCV polypeptide method for processing with compound provided herein.

Summary of the invention

In many embodiments, the invention provides the novel HCV proteinase inhibitor of a class, comprise one or more described compounds pharmaceutical composition, comprise one or more described compounds pharmaceutical preparation the preparation method and use one or more described compounds or one or more described preparation for treating or prevent HCV or improve the method for one or more symptoms of hepatitis C.HCV polypeptide and the interactional method of HCV proteolytic enzyme of regulating also is provided.In the compound provided by the invention, preferably suppress the compound of HCV NS3/NS4a serine protease.The invention discloses compound with following general formula I structure:

Formula I

Wherein:

Z be selected from heterocyclic radical ,-N (H) (alkyl) ,-N (alkyl) ₂,-N (H) (cycloalkyl) ,-N (cycloalkyl) ₂,-N (H) (aryl) ,-N (aryl) ₂,-N (H) (heterocyclic radical) ,-N (heterocyclic radical) ₂,-N (H) (heteroaryl) and-N (heteroaryl) ₂

R ¹Be H, OR ⁸, NR ⁹R ¹⁰Or CHR ⁹R ¹⁰, R wherein ⁸, R ⁹And R ¹⁰Can be identical or different, and independently be selected from separately H, alkyl-, thiazolinyl-, alkynyl-, aryl-, assorted alkyl-, heteroaryl-, cycloalkyl-, heterocyclic radical-, arylalkyl-and heteroarylalkyl, perhaps NR ⁹R ¹⁰In R ⁹And R ¹⁰Interconnect, make NR ⁹R ¹⁰Constitute 4-8 unit heterocyclic radical, same, CHR ⁹R ¹⁰In R ⁹And R ¹⁰Also can interconnect, make CHR ⁹R ¹⁰Constitute 4-8 unit cycloalkyl;

R ²And R ³Can be identical or different, and independently be selected from H, alkyl, assorted alkyl, thiazolinyl, assorted thiazolinyl, alkynyl, assorted alkynyl, cycloalkyl, heterocyclic radical, aryl, arylalkyl, heteroaryl and heteroarylalkyl separately;

Y is selected from following part:

Wherein G is NH or O; R ¹⁵, R ¹⁶, R ¹⁷, R ¹⁸, R ¹⁹, R ²⁰And R ²¹Can be identical or different, and independently be selected from H, alkyl, assorted alkyl, thiazolinyl, assorted thiazolinyl, alkynyl, assorted alkynyl, cycloalkyl, heterocyclic radical, aryl, arylalkyl, heteroaryl and heteroarylalkyl, perhaps (i) R separately ¹⁷And R ¹⁸Connect and compose 3-8 unit's cycloalkyl or heterocyclic radical independently of one another; (ii) R ¹⁵And R ¹⁹Connect and compose 4-8 unit heterocyclic radical independently of one another; (iii) R ¹⁵And R ¹⁶Connect and compose 4-8 unit heterocyclic radical independently of one another; (iv) R ¹⁵And R ²⁰Connect and compose 4-8 unit heterocyclic radical independently of one another;

Each described alkyl wherein; aryl; heteroaryl; cycloalkyl or heterocyclic radical can be unsubstitutedly or optional to be selected from following part and to replace by one or more independently: hydroxyl; alkoxyl group; aryloxy; sulfo-; alkylthio; arylthio; amino; amido; alkylamino; arylamino; alkyl sulphonyl; aryl sulfonyl; sulfonamido; alkyl; aryl; heteroaryl; amino-alkyl sulfinyl; aromatic yl sodium sulfonamido; oxo; carboxyl; carbalkoxy; formamido-; alkoxycarbonyl amino; alkoxyl group carbonyl oxygen base; the alkyl urea groups; aryl-ureido; halogen; cyano group and nitro.

In above-mentioned definition, the alkyl of preferred 1-10 carbon atom, the alkenyl or alkynyl of preferred 2-10 carbon atom, the preferred cycloalkyl of 3-8 carbon atom preferably has assorted alkyl, heteroaryl or the Heterocyclylalkyl (heterocyclic radical) of 1-6 oxygen, nitrogen, sulphur or phosphorus atom.

Formula I compound itself or unite one or more other suitable drug disclosed herein and can be used for treating multiple disease, for example HCV, HIV, AIDS (acquired immune deficiency syndrome (AIDS)) and relative disease, one or more symptoms that also can be used for regulating hepatitis C virus (HCV) protease activity, prevention HCV or improve hepatitis C.Both The compounds of this invention be can use, also can described adjusting, treatment, prevention or improvement be finished with pharmaceutical composition that comprises this compounds or preparation.Although be not limited to theory, believe that HCV proteolytic enzyme may be NS3 or NS4a proteolytic enzyme.The compounds of this invention can suppress this proteinoid enzyme.They can also regulate the processing of hepatitis C virus (HCV) polypeptide.

Detailed Description Of The Invention

In one embodiment, the invention discloses compound or its pharmacy acceptable salt or the solvate of structural formula I representative, wherein various piece as above defines.

In another embodiment, there is following stereoisomeric forms in any ratio in formula I compound:

Wherein various piece is suc as formula defining among the I.

In another embodiment, Z is selected from following part:

-N (methyl) ₂,

K=0-4 wherein, m=0-4, k and m can be identical or different, R ²⁶And R ²⁷Can be identical or different, and independently be selected from hydroxyl, alkoxyl group, aryloxy, sulfo-, alkylthio, arylthio, amino, amido, alkylamino, arylamino, alkyl sulphonyl, aryl sulfonyl, sulfonamido, alkyl, aryl, heteroaryl, amino-alkyl sulfinyl, aromatic yl sodium sulfonamido, oxo, carboxyl, carbalkoxy, formamido-, alkoxycarbonyl amino, alkoxyl group carbonyl oxygen base, alkyl urea groups, aryl-ureido, halogen, cyano group and nitro separately.

In another embodiment, R ¹Be NR ⁹R ¹⁰, R ⁹Be H, R ¹⁰Be H or R ¹⁴, R wherein ¹⁴Be alkyl, aryl, assorted alkyl, heteroaryl, cycloalkyl, alkyl-aryl, alkyl-heteroaryl, aryl-alkyl, thiazolinyl, alkynyl or heteroaryl-alkyl.

In another embodiment, R ¹⁴Be selected from following part:

With

In another embodiment, R ²Be selected from following part:

With

In another embodiment, R ³Be selected from following part:

R wherein ³¹Be OH or O-alkyl; R ³²Be H, C (O) CH ₃, C (O) O ^tBu or C (O) N (H) ^tBu.

In another embodiment, R ³Be selected from following part:

With

In another embodiment, G is NH.

In another embodiment, Y is selected from following part:

R wherein ¹⁵, R ¹⁶, R ¹⁷, R ¹⁸, R ¹⁹, R ²⁰And R ²¹Independently be selected from H, alkyl, assorted alkyl, thiazolinyl, assorted thiazolinyl, alkynyl, assorted alkynyl, cycloalkyl, heterocyclic radical, aryl, arylalkyl, heteroaryl and heteroarylalkyl, perhaps (i) R separately ¹⁷And R ¹⁸Connect and compose 3-8 unit's cycloalkyl or heterocyclic radical independently of one another; (ii) R ¹⁵And R ¹⁹Connect and compose 4-8 unit heterocyclic radical independently of one another; (iii) R ¹⁵And R ¹⁶Connect and compose 4-8 unit heterocyclic radical independently of one another; (iv) R ¹⁵And R ²⁰Connect and compose 4-8 unit heterocyclic radical independently of one another;

In another embodiment, with the lower section:

Be selected from:

With

Y wherein ³²Be selected from following groups:

With

In another embodiment, Y is selected from:

With

In another embodiment, Z is selected from the part with following structure:

-N (methyl) ₂,

In another embodiment, Z is selected from following part:

In another embodiment, Z is selected from following part:

In another embodiment, R ¹Be NH ₂Or NHR ¹⁴, R wherein ¹⁴Be selected from following part:

With

R ²Be selected from following part:

With

R ³Be selected from following part:

With

Z is selected from:

With

Y is selected from:

Another embodiment of the present invention discloses the cited compound of table 1.

Table 1

When following term is used, except as otherwise noted, otherwise be to be understood that to have following implication in context:

" patient " comprises humans and animals.

" Mammals " is meant people and other Mammals.

" alkyl " be meant and can be the aliphatic hydrocarbyl of straight or branched, and comprise in the chain about 1 to about 20 carbon atoms.Preferred alkyl in chain, comprise about 1 to about 12 carbon atoms.Preferred alkyl in chain, comprise about 1 to about 6 carbon atoms.Side chain is meant that one or more lower alkyls (for example methyl, ethyl or propyl group) are connected to linear alkyl chain." low alkyl group " is meant to have about 1 straight or branched group to about 6 carbon atoms in the chain.Term " alkyl of replacement " is meant the alkyl that is replaced by one or more identical or different substituting groups, each substituting group independently be selected from halogen, alkyl, aryl, cycloalkyl, cyano group, hydroxyl, alkoxyl group, alkylthio, amino ,-NH (alkyl) ,-NH (cycloalkyl) ,-N (alkyl) ₂,-N (alkyl) ₂, carboxyl and-C (O) O-alkyl.The limiting examples of suitable alkyl comprises methyl, ethyl, n-propyl, sec.-propyl and the tertiary butyl.

" thiazolinyl " is meant the aliphatic hydrocarbyl that contains at least one carbon-carbon double bond, it can be straight or branched and in chain, contain about 2 to about 15 carbon atoms.Preferred thiazolinyl in chain, contain about 2 to about 12 carbon atoms; More preferably in chain, contain about 2 to about 6 carbon atoms.Side chain is meant that one or more low alkyl groups (for example methyl, ethyl or propyl group) are connected to linear alkenylene chain." low-grade alkenyl " is meant and contains about 2 straight or branched thiazolinyls to about 6 carbon atoms in the chain.Term " thiazolinyl of replacement " is meant the thiazolinyl that is replaced by one or more identical or different substituting groups, each substituting group independently be selected from halogen, alkyl, aryl, cycloalkyl, cyano group, alkoxyl group and-S (alkyl).The limiting examples of suitable thiazolinyl comprises vinyl, propenyl, n-butene base, 3-methyl but-2-ene base, positive pentenyl, octenyl and decene base.

" alkynyl " is meant and contains at least one carbon carbon triple-linked aliphatic hydrocarbyl, it can be straight or branched and in chain, contain about 2 to about 15 carbon atoms.Preferred alkynyl in chain, contain about 2 to about 12 carbon atoms; More preferably in chain, contain about 2 to about 4 carbon atoms.Side chain is meant that one or more low alkyl groups (for example methyl, ethyl or propyl group) are connected to linear alkynyl chain." low-grade alkynyl " is meant and contains about 2 straight or branched alkynyls to about 6 carbon atoms in the chain.The limiting examples of suitable alkynyl comprises ethynyl, proyl, 2-butyne base and 3-methyl butynyl.Term " alkynyl of replacement " is meant the alkynyl that is replaced by one or more identical or different substituting groups, and each substituting group independently is selected from alkyl, aryl and cycloalkyl.

" aryl " be meant contain about 6 to about 14 carbon atoms, preferred about 6 to the aromatic monocyclic of about 10 carbon atoms or encircle ring system more.Aryl can be chosen wantonly by one or more identical or different " ring system substituting group " and replace, and the ring system substituting group is as definition herein.The limiting examples of suitable aryl comprises phenyl and naphthyl.

" heteroaryl " be meant contain about 5 to about 14 annular atomses, preferred about 5 to the aromatic monocyclic of about 10 annular atomses or encircle ring system more, wherein one or more annular atomses are not carbon atoms, for example are independent nitrogen, oxygen, sulphur atom or their combination.Preferred heteroaryl comprise about 5 to about 6 annular atomses." heteroaryl " can be chosen wantonly by one or more identical or different " ring system substituting group " and replace, and the ring system substituting group is as definition herein.Prefix azepine, oxa-or thia before the heteroaryl root name claims are meant that respectively at least one nitrogen, oxygen or sulphur atom are annular atoms.The nitrogen-atoms of heteroaryl can be chosen wantonly and be oxidized to the corresponding N oxide compound.The limiting examples of suitable heteroaryl comprises pyridyl, pyrazinyl, furyl, thienyl, pyrimidyl, pyridone (comprising the pyridone that N replaces), different _ the azoles base, isothiazolyl, _ azoles base, thiazolyl, pyrazolyl, the furazan base, pyrryl, pyrazolyl, triazolyl, 1,2, the 4-thiadiazolyl group, pyrazinyl, pyridazinyl, quinoxalinyl, 2,3 phthalazinyls, the oxindole base, imidazo [1,2-a] pyridyl, imidazo [2,1-b] thiazolyl, benzo furazan base, indyl, azaindolyl, benzimidazolyl-, benzothienyl, quinolyl, imidazolyl, the thienopyridine base, quinazolyl, the Thienopyrimidine base, pyrrolopyridinyl, imidazopyridyl, isoquinolyl, the benzo-aza indyl, 1,2, the 4-triazinyl, benzothiazolyl etc.Term " heteroaryl " also finger divides saturated heteroaryl moieties, for example tetrahydro isoquinolyl, tetrahydric quinoline group etc.

" aralkyl " or " arylalkyl " be meant aryl-alkyl-, wherein aryl and alkyl as above define.Preferred aralkyl comprises low alkyl group.The limiting examples of suitable aralkyl comprises benzyl, 2-styroyl and naphthyl methyl.Be connected to parent fraction by alkyl.

" alkylaryl " be meant alkyl-aryl-, wherein alkyl and aryl as above define.Preferred alkylaryl comprises low alkyl group.The limiting examples of suitable alkylaryl is a tolyl.Be connected to parent fraction by aryl.

" cycloalkyl " be meant the monocycle of non-aromatics or encircle ring system more, comprise about 3 to about 10 carbon atoms, preferred about 5 to about 10 carbon atoms.Preferred cycloalkyl ring comprise about 5 to about 7 annular atomses.Cycloalkyl can be chosen wantonly by one or more identical or different " ring system substituting group " and replace, and the ring system substituting group is as definition herein.The limiting examples of suitable monocyclic cycloalkyl comprises cyclopropyl, cyclopentyl, cyclohexyl, suberyl etc.The limiting examples of suitable polycyclic naphthene base comprises 1-decahydro naphthyl, norborneol alkyl, adamantyl etc., also comprises the polycyclic naphthene base of fractional saturation, for example indanyl, tetralyl etc.

" halogen " is meant fluorine, chlorine, bromine or iodine.Preferred fluorine, chlorine and bromine.

" ring system substituting group " is meant the substituting group that is connected to aromatics or non-aromatics ring system, for example, and the available hydrogen on its replaceable described ring system.Each ring system substituting group can be identical or different, and independently be selected from following group separately: alkyl; thiazolinyl; alkynyl; aryl; heteroaryl; aralkyl; alkylaryl; heteroaralkyl; the heteroaryl thiazolinyl; the heteroaryl alkynyl; miscellaneous alkyl aryl; hydroxyl; hydroxyalkyl; alkoxyl group; aryloxy; aralkoxy; acyl group; aroyl; halogen; nitro; cyano group; carboxyl; alkoxy carbonyl; aryloxycarbonyl; aromatic alkoxy carbonyl; alkyl sulphonyl; aryl sulfonyl; heteroarylsulfonyl; alkylthio; arylthio; heteroarylthio; aromatic alkyl sulfurio; the heteroaralkyl sulfenyl; cycloalkyl; heterocyclic radical;-C (=N-CN)-NH ₂,-C (=NH)-NH ₂,-C (=NH)-NH (alkyl), Y ₁Y ₂N-, Y ₁Y ₂The N-alkyl-, Y ₁Y ₂NC (O)-, Y ₁Y ₂NSO ₂-and-SO ₂NY ₁Y ₂, Y wherein ₁And Y ₂Can be identical or different, and independently be selected from hydrogen, alkyl, aryl, cycloalkyl and aralkyl." ring system substituting group " can refer to that also D-loop simultaneously fastens a part of two available hydrogens (H on each carbon) of two adjacent carbonss.The substituent example of this class ring system have methylene-dioxy, ethylenedioxy ,-C (CH ₃) ₂-etc., their configuration examples such as following part:

With

" heterocyclic radical " is meant monocycle or many ring fillings ring system of non-aromatics, comprise about 3 to about 10 carbon atoms, preferred about 5 to about 10 carbon atoms, wherein one or more annular atomses are not carbon atoms, for example are independent nitrogen, oxygen, sulphur atom or their combination.There are not adjacent oxygen and/or sulphur atom in the ring system.Preferred heterocyclic radical comprise about 5 to about 6 annular atomses.Prefix azepine, oxa-or thia before the heterocyclic radical root name claims are meant that respectively at least one nitrogen, oxygen or sulphur atom are annular atoms.Any in the heterocyclic ring-NH can be protected form, for example-N (Boc) ,-N (CBz) ,-N (Tos) etc.; Such protection also is considered to integral part of the present invention.Heterocyclic radical can be chosen wantonly by one or more identical or different " ring system substituting group " and replace, and the ring system substituting group is as definition herein.The nitrogen of heterocyclic radical or sulphur atom can be chosen wantonly and be oxidized to corresponding N-oxide compound, S-oxide compound or S, the S-dioxide.The limiting examples of suitable monocyclic heterocycles base comprises piperidyl, pyrrolidyl, piperazinyl, morpholinyl, thio-morpholinyl, thiazolidyl, 1,4-two _ alkyl, tetrahydrofuran base, tetrahydro-thienyl, lactan, lactone etc.

Should be noted in the discussion above that in the present invention to contain in the heteroatomic ring system on the carbon atom adjacent, do not have hydroxyl, on the carbon atom adjacent, also do not have N or S group with another heteroatoms with N, O or S.Therefore, for example in following ring:

Do not have-OH directly is connected on 2 and 5 s' the carbon atom.

Should also be noted that tautomeric forms, for example following part:

With

In certain embodiments of the invention, be considered to be equal to.

" alkynyl alkyl " be meant alkynyl-alkyl-, wherein alkynyl and alkyl as above define.Preferred alkynyl alkyl comprises low-grade alkynyl and low alkyl group.Be connected to parent fraction by alkyl.The limiting examples of suitable alkynyl alkyl comprises the propargyl methyl.

" heteroaralkyl " be meant heteroaryl-alkyl-, wherein heteroaryl and alkyl as above define.Preferred heteroaralkyl comprises low alkyl group.The limiting examples of suitable aralkyl comprises pyridylmethyl and quinoline-3-ylmethyl.Be connected to parent fraction by alkyl.

" hydroxyalkyl " be meant the HO-alkyl-, wherein alkyl as above defines.Preferred hydroxyalkyl comprises low alkyl group.The limiting examples of suitable hydroxyalkyl comprises hydroxymethyl and 2-hydroxyethyl.

" acyl group " be meant H-C (O)-, alkyl-C (O)-or cycloalkyl-C (O)-, alkyl and cycloalkyl as above define.Be connected to parent fraction by carbonyl.Preferred acyl group comprises low alkyl group.The limiting examples of suitable acyl group comprises formyl radical, ethanoyl and propionyl.

" aroyl " be meant aryl-C (O)-, wherein aryl as above defines.Be connected to parent fraction by carbonyl.The limiting examples of suitable aroyl comprises benzoyl and 1-naphthoyl.

" alkoxyl group " is meant alkyl-O-, and wherein alkyl as above defines.The limiting examples of suitable alkoxyl group comprises methoxyl group, oxyethyl group, positive propoxy, isopropoxy and n-butoxy.Be connected to parent fraction by ether oxygen.

" aryloxy " is meant aryl-O-, and wherein aryl as above defines.The limiting examples of suitable aryloxy comprises phenoxy group and naphthyloxy.Be connected to parent fraction by ether oxygen.

" aralkyl oxy " is meant aralkyl-O-, and wherein aralkyl as above defines.The limiting examples of suitable aralkyl oxy comprises benzyloxy and 1-naphthalene methoxyl group or 2-naphthalene methoxyl group.Be connected to parent fraction by ether oxygen.

" alkylthio " is meant alkyl-S-, and wherein alkyl as above defines.The limiting examples of suitable alkylthio comprises methylthio group and ethylmercapto group.Be connected to parent fraction by sulphur.

" arylthio " is meant aryl-S-, and wherein aryl as above defines.The limiting examples of suitable arylthio comprises thiophenyl and naphthalene sulfenyl.Be connected to parent fraction by sulphur.

" aromatic alkyl sulfurio " is meant aralkyl-S-, and wherein aralkyl as above defines.The limiting examples of suitable aromatic alkyl sulfurio is a benzylthio-.Be connected to parent fraction by sulphur.

" alkoxy carbonyl " is meant alkyl-O-CO-.The limiting examples of suitable alkoxy carbonyl comprises methoxycarbonyl and ethoxy carbonyl.Be connected to parent fraction by carbonyl.

" aryloxycarbonyl " be meant aryl-O-C (O)-.The limiting examples of suitable aryloxycarbonyl comprises phenyloxycarbonyl and naphthyloxy carbonyl.Be connected to parent fraction by carbonyl.

" aromatic alkoxy carbonyl " be meant aralkyl-O-C (O)-.The limiting examples of suitable aromatic alkoxy carbonyl is a benzyloxycarbonyl.Be connected to parent fraction by carbonyl.

" alkyl sulphonyl " is meant alkyl-S (O ₂)-.Alkyl is a low alkyl group in the preferred alkyl sulphonyl.Be connected to parent fraction by alkylsulfonyl.

" aryl sulfonyl " is meant aryl-S (O ₂)-.Be connected to parent fraction by alkylsulfonyl.

Term " replace () " being meant that the one or more hydrogen on specified atom are replaced by selected appointment group, precondition is the normal valency that is no more than specified atom under the existing situation, and this replacement obtains stable compound.The combination of substituting group and/or variable only just is allowed under this combination obtains the situation of stable compound." stable compound " or " rock steady structure " is meant that compound is enough firm, can from reaction mixture, separate reach can be practical purity, and can be formulated as effective medicine.

When term " one or more " or " at least a " are used to refer to the quantity of substituting group, compound, combination medicine etc., be meant existence or add at least a substituting group, compound, combination medicine etc., the maximum quantity that can reach chemistry at most and physically allow, concrete quantity depends on context.This class technology and knowledge are well known to a person skilled in the art.

Term " optional replacement " is meant that optional designated group or part replace.

The term of allied compound " isolating " or " unpack format () " be meant the physical condition after described compound separates down from building-up process, natural origin or both of these case.The term of allied compound " purifying () " or " purified form () " be meant that described compound is through physical condition this paper introduction or that well known to a person skilled in the art one or more purge processes acquisitions, and have enough purity, so as by this paper introduction or well known to a person skilled in the art that standard analytical techniques characterizes.

Should also be noted that and in the application's text, flow process, embodiment and form, do not satisfy valent any carbon atom or heteroatoms satisfies valency by hydrogen atom.

When certain functional group was " protection " in the compound, this was meant that this group is a modified forms, thus when compound reacts, prevent to take place in protected position unwanted pair sends out should.Suitable protecting group is that those of ordinary skills know, but also reference standard textbook, T.W.Greene etc. for example, Protective Groups in organic Synthesis (1991), Wiley, New York.

Any variable (for example aryl, heterocycle, R ²Deng) when occurring surpassing one time in any composition or formula I, the variable-definition in each position is independent of the variable-definition of all other positions.

Term used herein " composition " comprises the product of the predetermined component that contains prescribed dose and is made up directly or indirectly and any product of obtaining by the predetermined component of prescribed dose.

The prodrug and the solvate of The compounds of this invention are also included within the scope of the invention.Term used herein " prodrug " is meant the compound into prodrug, in case after giving the patient, through metabolic process or chemical process generation chemical conversion, obtain formula I compound or its salt and/or solvate.The elaboration of prodrug is referring to T.Higuchi and V.Stella, Pro-drugs asNovel Delivery Systems (1987), A.C.S.Symposium Series, the 14th volume; Bioreversible Carriers in Drug Design, (1987) Edward B.Roche edits, American Pharmaceutical Association and Pergamon Press, these two pieces of documents all are attached to herein by reference.

" solvate " is meant the The compounds of this invention with one or more solvent molecule physical bond.This physical bond relates to ionic linkage combination and covalent bonds in various degree, comprises hydrogen bonded.In some cases, solvate can be separated, for example when one or more solvent molecules are attached to the lattice of crystalline solid." solvate " comprises solution phase and separable solvate.The limiting examples of suitable solvent compound comprises ethanol compound, methyl alcohol compound etc." hydrate " is that solvent molecule is H ₂The solvate of O.

" significant quantity " or " treatment significant quantity " is meant that the consumption of The compounds of this invention or composition can effectively suppress serine protease, and obtains required treatment, improvement, inhibition or preventive effect thus.

Formula I compound can form the salt that also belongs to the scope of the invention.Except as otherwise noted, otherwise should be understood to comprise its salt when mentioning formula I compound.Term used herein " salt " is meant acid salt that forms with mineral acid and/or organic acid and the subsalt that forms with mineral alkali and/or organic bases.In addition, when formula I compound not only comprises basic moiety (such as but not limited to pyridine or imidazoles) but also comprises acidic moiety (such as but not limited to carboxylic acid), can form zwitter-ion (" inner salt "), zwitter-ion is also included within the term used herein " salt ".Although other salt also is useful, preferably pharmaceutically acceptable (promptly nontoxic, physiologically acceptable) salt.The salt of formula I compound can generate by for example following mode: make formula I compound and a certain amount of acid or alkali (for example 1 equivalent) reaction, medium uses the sedimentary therein medium of for example described salt, perhaps uses aqueous medium and with postlyophilization.

Exemplary acid salt comprise acetate, ascorbate salt, benzoate, benzene sulfonate, hydrosulfate, borate, butyrates, Citrate trianion, camphorate, camsilate, fumarate, hydrochloride, hydrobromate, hydriodate, lactic acid salt, maleate, mesylate, naphthalenesulfonate, nitrate, oxalate, phosphoric acid salt, propionic salt, salicylate, succinate, vitriol, tartrate, thiocyanate-, tosylate etc.In addition, it has been generally acknowledged that to be fit to generate in the document of acid below for example of pharmaceutical salts and be described: P.Stahl etc. with the basic medicinally compound, Camille G. (editor) Handbook ofPharmaceutical Salts.Properties, Selection and Use. (2002) Zurich:Wiley-VCH; S.Berge etc., Journal of Pharmaceutical Sciences (1977) 66 (1) 1-19; P.Gould, International J.of Pharmaceutics (1986) 33201-217; Anderson etc., The Practice of Medicinal Chemistry (1996), AcademicPress, New York; The Orange Book is (at Food ﹠amp; Drug Administration, Washington is on the D.C. website).The disclosure of above-mentioned document is attached to herein by reference.

Exemplary basic salts comprises ammonium salt, an alkali metal salt (for example sodium salt, lithium salts and sylvite), alkaline earth salt (for example calcium salt and magnesium salts), the salt that forms with organic bases (for example organic amine, for example dicyclohexylamine, TERTIARY BUTYL AMINE) and the salt that forms with amino acid (for example arginine, Methionin etc.).The alkalescence nitrogen-containing group can be with for example following reagent is quaternized: low alkyl group halogen (for example muriate of methyl, ethyl and butyl, bromide and iodide), sulfuric acid dialkyl (for example methyl-sulfate, sulfuric acid diethyl ester and dibutyl sulfate), long-chain halogenide (for example muriate of decyl, dodecyl and stearyl, bromide and iodide), aralkyl halogen (for example bromotoluene and phenethyl bromide) and quaternized with other reagent.

Acid salt that all are such and subsalt all are the pharmacy acceptable salts in the scope of the invention, and think that for the object of the invention all acid salt and subsalt all are equal to the free form of respective compound.

May there be (for example being acid amides or imino-ether) with their tautomeric forms in formula I compound and salt thereof, solvate and prodrug.All these class tautomeric forms all are integral parts of the present invention.

All steric isomers (for example geometrical isomer, optically active isomer etc.) of The compounds of this invention (comprising salt, solvate and the prodrug of The compounds of this invention and the salt and the solvate of described prodrug) are included in the scope of the present invention, the steric isomer that exists owing to the asymmetric carbon of different substituents for example, comprise enantiomeric forms (even not having asymmetric carbon also may exist), rotational isomer form, atropisomer and diastereomeric form, also comprise positional isomers (for example 4-pyridyl and 3-pyridyl).Each steric isomer of The compounds of this invention can be the form that does not for example have other isomer substantially, perhaps can be mixed into for example racemoid, perhaps mixes with all other steric isomers or selected other steric isomer.Chiral centre of the present invention can have the S or the R configuration of IUPAC 1974 Recommendations definition.When terms such as use " salt ", " solvate ", " prodrug ", be equally applicable to salt, solvate and the prodrug of enantiomer, steric isomer, rotational isomer, tautomer, positional isomers, racemic modification or the prodrug of The compounds of this invention.

The present invention also comprises the polymorphic form of formula I compound and salt, solvate and prodrug.

Should be understood that the formula I application of compound that is used for therepic use described herein is applicable to each compound itself, perhaps one or more combinations of one or more formulas I compound are as the explanation of next section example.Same understanding also is applicable to pharmaceutical composition that contains this compounds or the methods of treatment that relates to this compounds.

The compounds of this invention can have pharmacological characteristics; The special useful as HCV proteinase inhibitor of formula I compound, each compound itself or one or more formulas I compound can make up with one or more compounds that are selected from formula I.The compounds of this invention can be used for treating multiple disease, for example HCV, HIV (AIDS, acquired immune deficiency syndrome (AIDS)) and relative disease, also can regulate hepatitis C virus (HCV) protease activity, prevent HCV or improve one or more symptoms of hepatitis C.

Formula I compound can be used for preparing the medicine that is used for the treatment of HCV proteolytic enzyme relative disease, and for example described preparation method comprises makes formula I compound closely contact with pharmaceutically acceptable carrier.

In another embodiment, the invention provides pharmaceutical composition, said composition comprises one or more The compounds of this invention as activeconstituents.Usually, pharmaceutical composition also comprises at least a pharmaceutically acceptable carrier thinner, vehicle or carrier (the present invention is referred to as solid support material).They because having HCV, this class pharmaceutical composition suppress active, so can be used for treating hepatitis C and relative disease thereof.

In another embodiment, the invention discloses preparation and comprise the method for The compounds of this invention as the pharmaceutical composition of activeconstituents.In pharmaceutical composition of the present invention and method, activeconstituents usually with the suitable carriers material mixing after administration, form of medication is as required suitably selected solid support material, be oral tablet, capsule (solid is filled, the semi-solid filling or liquid filling) but, reprovision is with powder, oral gel, elixir dispersible granule, syrup, suspensoid etc., and meets the conventional pharmaceutical practice.For instance, for with tablet or Capsule form oral administration, active pharmaceutical ingredient can mix with any oral nontoxic pharmaceutically acceptable inert support, for example lactose, starch, sucrose, Mierocrystalline cellulose, Magnesium Stearate, Lin Suanergai, calcium sulfate, talcum powder, mannitol, ethanol (liquid form) etc.In addition, when requiring or need, also suitable binder, lubricant, disintegrating agent and tinting material can be admixed in the mixture.Powder and tablet can comprise about 5% to about 95% component of the present invention.

Suitable binder comprises starch, gelatin, natural sugar, corn sweetener, natural and synthetic gum (as Sudan Gum-arabic), sodiun alginate, carboxymethyl cellulose, polyoxyethylene glycol and wax.The examples of lubricant that can be used for above-mentioned formulation has boric acid, Sodium Benzoate, sodium acetate, sodium-chlor etc.Disintegrating agent comprises starch, methylcellulose gum, guar gum etc.

Also can comprise sweeting agent, seasonings and sanitas under the suitable situation.Set forth more above-mentioned terms hereinafter in more detail, i.e. disintegrating agent, thinner, lubricant, tackiness agent etc.

In addition, composition of the present invention can be mixed with sustained release form, thereby the rate of release of controlling any or multiple composition or activeconstituents is to reach optimized result of treatment, and promptly HCV suppresses activity etc.The dosage forms that continues to discharge comprises multilayer tablet, it comprises the layer or the controlled release polymer matrix of a plurality of different disintegration rate, activeconstituents is immersed in the polymeric matrix, is processed into tablet form, perhaps comprises the capsule of described dipping or parcel porous polymer matrix.

Liquid preparation comprises solution, suspensoid and emulsion.For example be used for the aqueous pharmaceutical of parenteral injection or oral solution, suspensoid and the emulsion of water-propylene glycol solution agent or adding sweeting agent and opalizer (pacifiers).Liquid preparation also can comprise the intranasal administration solution.

The aerosol formulations that is suitable for sucking can comprise the solid of solution and powder type, they can with pharmaceutically acceptable carrier inertia pressurized gas (as nitrogen) combined utilization for example.

Preparation is during suppository, the low-melting wax of fusion at first, the mixture of glycerin fatty acid ester (as theobroma oil) for example, and by stir or similarly blending means activeconstituents is dispersed in wherein.Then the fused uniform mixture is poured in the mould of suitable size, made it cooling, solidify thus.

Also comprise such solid preparation: can face with before being mixed with oral or the administered parenterally liquid preparation.Described liquid preparation comprises solution, suspensoid and emulsion.

But The compounds of this invention is transdermal administration also.The percutaneous administration composition can adopt ointment, lotion, aerosol and/or emulsion form, and described composition can be included in matrix type or the reservoir devices transdermal patch, and this is to be used for this purpose routine techniques in this area.

The compounds of this invention also can be oral, in the intravenously, sheath, in the nose or subcutaneous administration.

The compounds of this invention also comprises the preparation of unit dosage.In this class formulation, preparation is subdivided into the unitary dose of the suitable size that comprises an amount of (for example reaching the significant quantity of required purpose) activeconstituents.

According to concrete application, the dosage of activeconstituents of the present invention is normally adjustable in the unit dose formulations, about 1.0 milligrams to about 1,000 milligram, preferred about 1.0 milligrams to about 950 milligrams, more preferably from about 1.0 milligrams to about 500 milligrams, typical doses is about 1 milligram to about 250 milligrams.Actual using dosage can change according to patient's age, sex, body weight and the degree of being in a bad way of being treated.This technology is that those skilled in the art are well-known.

Usually, can give to comprise for 1 time or 2 times the human oral dosage form of activeconstituents every day.Dosage and administration frequency need be adjusted according to attending doctor's judgement.The general recommended scope of oral administration from about 1.0 mg/day to about 1,000 mg/day, can be once or the gradation administration.

Some useful terms are described as follows:

Capsule is meant special container or encapsulation object, and they are made by methylcellulose gum, polyvinyl alcohol or metagelatin or starch, is used for keeping or holding comprising composition of active components.Hard-shell capsule is generally made by the adulterant of high-intensity relatively gelatin skeleton and pigskin gelatin.Capsule itself can comprise a small amount of dyestuff, opacifying agent, softening agent and sanitas.

Tablet is meant compacting or the molded solid formulation that comprises activeconstituents and suitable diluents.By compressing mixt or the granules preparation tablet that obtains through wet granulation, dry granulation or compression.

Oral gel is meant that activeconstituents is scattered in or is dissolved in the wetting ability semisolid matrix.

Reprovision is meant the powder adulterant that comprises activeconstituents and suitable diluents with powder, its available being suspended in water or the fruit juice.

Thinner is meant the material of common formation composition or formulation major portion.Suitable diluent comprises carbohydrate, for example lactose, sucrose, mannitol and Sorbitol Powder; Starch from wheat, corn, rice and potato acquisition; Mierocrystalline cellulose, for example Microcrystalline Cellulose.The weight percent that amount of diluent accounts for total composition in the composition from about 10% to about 90%, preferred about 25% is to about 75%, and more preferably from about 30% to about 60%, and more more preferably from about 12% to about 60%.

Disintegrating agent is meant and joins in the composition so that the material that impels it to separate (disintegration) and discharge medicine.Suitable disintegrants comprises starch; " coldwater-soluble " treated starch, for example sodium starch glycolate; Natural and synthetic gum class is as tracasol, karaya, guar gum, tragacanth gum and agar; Derivatived cellulose is as methylcellulose gum and Xylo-Mucine; Microcrystalline Cellulose and crosslinked Microcrystalline Cellulose, for example croscarmellose sodium; Alginate, for example alginic acid and sodiun alginate; Clay, for example wilkinite; And effervescent mixture.The weight percent that disintegrant content accounts for composition in the composition is from about 2% to about 15%, and more preferably from about 4% to about 10%.

Tackiness agent is to instigate powder-stuck or " deadlocked " together and make their adhesive aggregations form the particulate material, therefore when preparation as " tackiness agent ".Tackiness agent increases thinner or the existing adhesive strength of extender.Suitable binder comprises carbohydrate such as sucrose; Starch from wheat, corn, rice and potato acquisition; Natural gum class, for example Sudan Gum-arabic, gelatin and tragacanth gum; Marine alga derivative, for example alginic acid, sodiun alginate and Protanal TXF 200 ammonium; Cellulose materials, for example methylcellulose gum, Xylo-Mucine and Vltra tears; Polyvinylpyrrolidone; And inorganics magnesium aluminum silicate for example.The weight percent that binder content can account for composition in the composition is from about 2% to about 20%, and more preferably from about 3% to about 10%, and more more preferably from about 3% to about 6%.

Lubricant is meant in the formulations such as joining tablet, granule, so as after compacting the material by reducing friction or wearing and tearing preparation is deviate from from mould or punch die.Examples of suitable lubricants comprises metallic stearate, for example Magnesium Stearate, calcium stearate or potassium stearate; Stearic acid; High melting-point wax; And soluble oil, for example sodium-chlor, Sodium Benzoate, sodium acetate, sodium oleate, polyoxyethylene glycol and d ' l-leucine.The lubricant final step before compacting usually adds, because they must be present in the particulate surface and between particle and tabletting machine assembly.Lubricant content can account for composition weight per-cent from about 0.2% to about 5% in the composition, and preferred about 0.5% to about 2%, and more preferably from about 0.3% to about 1.5%.

Glidant (glident)-prevent caking and improve particulate mobile so that the level and smooth material uniformly that flows.Suitable glidant comprises silicon-dioxide and talcum powder.Glidant content can account for the total weight percent from about 0.1% to about 5%, preferred about 0.5% to about 2% of composition in the composition.

Tinting material-make composition or the painted vehicle of formulation.This class vehicle can comprise food grade dyes and the food grade dyes that is adsorbed onto on the suitable adsorbent (for example clay or aluminum oxide).Colorant content can account for the weight percent from about 0.1% to about 5%, preferred about 0.1% to about 1% of composition.

Bioavailability is meant with standard substance or contrast and compares that the active pharmaceutical ingredient or the treatment of the formulation that gives are partially absorbed into body round-robin speed and degree.

The ordinary method of preparation tablet is that people are known.These class methods comprise dry method (for example directly the particle that compression produces is passed through in compacting and compacting), wet method or other special methods.The ordinary method for preparing other form of administration (for example capsule, suppository etc.) also is that people know.

Another embodiment of the invention discloses above disclosed The compounds of this invention or the pharmaceutical composition purposes in treatment disease (for example hepatitis C etc.).This method comprises the The compounds of this invention or the pharmaceutical composition of the patient treatment significant quantity of suffering from one or more described diseases and this treatment of needs.

In another embodiment, The compounds of this invention can be used for the treatment of human HCV, and its therapeutic modality can be monotherapy or conjoint therapy (for example dual associating, triple unite etc.), for example combined utilization antiviral drug and/or immunomodulator.The example of this class antiviral drug and/or immunomodulator comprise ribavirin (Schering-Plough Corporation, Madison, NewJersey) and Levovirin ^TM(ICN Pharmaceuticals, Costa Mesa, California), VP 50406 ^TM(Viropharma, Incorporated, Exton, Pennsylvania), ISIS14803 ^TM(ISIS Pharmaceuticals, Carlsbad, Califbrnia), Heptazyme ^TM(Ribozyme Pharmaceuticals, Boulder, Colorado), VX 497 ^TM(VertexPharmaceuticals, Cambridge, Massachusetts), Thymosin ^TM(SciClonePharmaceuticals, San Mateo, California), Maxamine ^TM(MaximPharmaceuticals, San Diego, California), mycophenolate mofetil (Hoffman-LaRoche, Nutley, New Jersey), Interferon, rabbit (for example interferon alpha, polyethylene glycol-interferon alpha conjugate) etc." polyethylene glycol-interferon alpha conjugate " is the interferon alpha molecule of covalently bound peg molecule.The example of polyethylene glycol-interferon alpha conjugate comprises that the Intederon Alpha-2a form that adds polyoxyethylene glycol is (for example with trade(brand)name Pegasys ^TMSale) Intederon Alpha-2a (Roferon ^TM, Hoffman La-Roche, Nutley, New Jersey), add the Interferon Alpha-2b form of polyoxyethylene glycol (for example with trade(brand)name PEG-Intron ^TMSale) Interferon Alpha-2b (Intron ^TM, Schering-Plough Corporation), interferon c (BeroforAlpha ^TM, Boehringer Ingelheim, Ingelheim, Germany) or the total Interferon, rabbit (Infergen of the consensus sequence definition by measuring natural interferon alpha ^TM, Amgen, Thousand Oaks, California).

As mentioned above, the present invention also comprises tautomer, rotational isomer, enantiomer and other steric isomer of The compounds of this invention.Therefore, those skilled in the art can be understood that compounds more of the present invention may exist with suitable isomeric forms.Such version also belongs to the scope of the invention.

Another embodiment of the invention discloses the preparation method of The compounds of this invention.Described compound can be by several technology preparations known in the art.Following reaction process summary description illustrative methods.These example explanations should not be interpreted as the restriction to the scope of the invention of appended claims definition.Approach that other is machine-processed and similar structures are conspicuous to those skilled in the art.

Should be understood that,, suitably replace any natural and alpha-non-natural amino acid and will form required compound based on described replacement though following exemplary flow has been introduced the preparation method of a small amount of representative compounds of the present invention.This class variation also belongs to the scope of the invention.

Abbreviation:

In the description of following flow process, preparation example and embodiment, use following abbreviation:

THF: tetrahydrofuran (THF)

DMF:N, dinethylformamide

EtOAc: ethyl acetate

AcOH: acetate

HOOBt:3-hydroxyl-1,2,3-phentriazine-4 (3H)-ketone

EDCI:1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride

The NMM:N-methylmorpholine

ADDP:1,1 '-(azo dicarbapentaborane (azodicarbobyl)) two piperidines

DEAD: diethyl azodiformate

MeOH: methyl alcohol

EtOH: ethanol

Et ₂O: ether

DMSO: methyl-sulphoxide

The HOBt:N-hydroxybenzotriazole

PyBrOP: phosphofluoric acid bromo-three-pyrrolidyl rattle

DCM: methylene dichloride

DCC:1, the 3-dicyclohexylcarbodiimide

TEMPO:2,2,6,6-tetramethyl piperidine-1-oxyradical

Phg: phenylglycocoll

Chg: Cyclohexylglycine

Bn: benzyl

Bzl: benzyl

Et: ethyl

Ph: phenyl

IBoc: isobutoxy carbonyl

IPr: sec.-propyl

^tBu or Bu ^t: the tertiary butyl

Boc: tert-butoxycarbonyl

Cbz: benzyloxycarbonyl

Cp: cyclopentadienyl

Ts: p-toluenesulfonyl

Me: methyl

HATU: phosphofluoric acid O-(7-azepine benzo triazol-1-yl)-1,1,3, the 3-tetramethyl-urea _

DMAP:4-N, the N-dimethyl aminopyridine

BOP: benzotriazole-1-base-oxygen-three (dimethylamino) hexafluorophosphate

PCC: pyridinium chlorochromate _

KHMDS: hexamethyl two silica-based azane potassium or two (trimethyl silicon based) potassium amide

NaHMDS: hexamethyl two silica-based azane sodium or two (trimethyl silicon based) sodium amide

LiHMDS: hexamethyl two silica-based azane lithiums or two (trimethyl silicon based) Lithamide

10%Pd/C:10% palladium on carbon (by weight).

TG: thioglycerin

Embodiment:

The preparation of intermediate:

Preparation intermediate 1.01:

Step 1:

To 4-oxyproline derivative 1.02 (1.0g, 4.1mmol) and in 0 ℃ of solution of methylene dichloride (50ml), add successively chloroformic acid 4-nitro phenyl ester (2.46g, 12.2mmol), pyridine (0.987ml, 12.2mmol).After keeping 15 minutes under this temperature, reaction flask is placed on spend the night in the refrigerator (20 ℃) (16 hours).Reaction mixture is with methylene dichloride (100ml) dilution, with saturated ammonium chloride solution (2 * 100ml), salt solution (100ml) washing, through dried over sodium sulfate, filter the back and concentrate.Crude product obtains 1.03 (1.5g) with silica gel chromatography purifying (20/80 to 50/50 EtOAc/ hexane).

Step 2:

To proline derivative 1.03 (1.5g 3.66mmol) and in 0 ℃ of stirred solution of methylene dichloride (30ml), adds 1,2,3, the 4-tetrahydroisoquinoline (0.55ml, 4.39mmol) and DIPEA (2.02ml, 10.98mmol).After keeping 15 minutes under this temperature, reaction flask is placed on spend the night in the refrigerator (20 ℃) (16 hours).Reaction mixture is with methylene dichloride (70ml) dilution, with saturated ammonium chloride solution (100ml), saturated sodium bicarbonate solution (3 * 100ml), salt solution (100ml) washing, through dried over sodium sulfate, filter the back and concentrate.Crude product obtains the required product 1.04 of 1.4g with silica gel chromatography purifying (5/95 to 25/75 EtOAc/ methylene dichloride).LC-MS：405.1(M+H) ⁺。

Step 3:

In the product 1.04 (1.4g) of above acquisition, add 4M HC1/ two _ alkane (25ml).Reactant was at room temperature placed 1 hour, concentrated then, obtain required intermediate 1.01, directly use to need not repurity with quantitative output.

Preparation intermediate 10.11 and 10.12:

Step 1:

Under nitrogen atmosphere, (50g, 187.1mmol) stirred solution with anhydrous THF (400ml) is cooled to-78 ℃, with 1M potassium tert.-butoxide (K-with ketoimine 10.01 ^tBuO) (220ml, THF solution-treated 1.15eq.).Reaction mixture is risen to 0 ℃, stirred 1 hour, (28ml 249mmol) handles with the brooethyl tetramethylene.Reaction mixture was at room temperature stirred 48 hours vacuum concentration.Resistates is dissolved in ether (300ml), and (2M 300ml) handles, and gained solution is at room temperature stirred 5 hours, with ether (1L) extraction with the HCl aqueous solution.Water layer alkalizes to pH～12-14 with NaOH (50% aqueous solution), uses CH ₂Cl ₂(3 * 300ml) extractions.The organic layer that merges filters the back and concentrates through dried over mgso, obtain the pure amine of colorless oil (10.02,18g).

Step 2

Amine 10.02 (18g, 105.2mmol) and CH ₂Cl ₂(23g 105.4mmol) handles 0 ℃ of solution (350ml), at room temperature stirs 12 hours with tert-Butyl dicarbonate.After complete reaction (TLC), the vacuum concentration reaction mixture is dissolved in THF/H with resistates ₂O (200ml, 1: 1) uses LiOHH ₂(6.5g 158.5mmol) handles O, at room temperature stirs 3 hours.Concentrated reaction mixture, alkaline water layer Et ₂The O extraction.Water layer is acidified to pH～1-2 with concentrated hydrochloric acid, uses dichloromethane extraction.The organic layer that merges filters final vacuum and concentrates through dried over mgso, obtains colourless viscosity oily matter 10.03, is directly used in next step and need not repurity.

Step 3

Acid 10.03 (15.0g, CH 62mmol) ₂Cl ₂(250ml) (41.1g, 93mmol), N-methylmorpholine (27ml), N, (9.07g 93mmol) handles the O-dimethyl hydroxylamine hydrochloride solution, at room temperature stirs and spends the night with bop reagent.Reaction mixture is isolated each layer, water layer CH with the 1N HCl aqueous solution (250ml) dilution ₂Cl ₂(3 * 300ml) extractions.The organic layer that merges filters through dried over mgso, and vacuum concentration is with chromatography purification (SiO ₂, EtOAc/ hexane 2: 3), obtain colorless solid acid amides 10.04 (15.0g).

Step 4

At 0 ℃, (15g drips LiAlH in anhydrous THF (200ml) solution 52.1mmol) to acid amides 10.04 ₄(1M, 93ml, 93mmol) solution.Reaction mixture was at room temperature stirred 1 hour, use KHSO at 0 ℃ ₄The careful quencher of solution (10% aqueous solution) was stirred 0.5 hour.(1M, 150ml) CH is used in dilution to reaction mixture with the HCl aqueous solution ₂Cl ₂(3 * 200ml) extractions, the organic layer of merging is with the HCl aqueous solution (1M), saturated sodium bicarbonate, salt water washing, through dried over mgso.Filtering mixt, vacuum concentration obtains colourless viscosity oily matter 10.05 (14g).

Step 5

Aldehyde 10.05 (14g, CH 61.6mmol) ₂Cl ₂(50ml) solution Et ₃N (10.73ml, 74.4mmol) and acetone cyanohydrin (10.86g 127.57mmol) handles, and at room temperature stirs 24 hours.The dense vacuum reaction mixture that contracts, (1M, 200ml) dilution is extracted into CH with the HCl aqueous solution ₂Cl ₂(in 3 * 200ml).The organic layer water, the salt water washing that merge through dried over mgso, are filtered, and vacuum concentration is with chromatography purification (SiO ₂, EtOAc/ hexane 1: 4), obtain colourless liquid 10.06 (10.3g).

Step 6

The saturated methanol solution of HCl ^*(by at 0 ℃ to CH ₃OH (700ml) feeds HCl gas and prepares) handle with cyanalcohol 10.06, be heated to and refluxed 24 hours.The vacuum concentration reactant obtains 10.07, is directly used in next step and need not repurity.

^*Also can use to anhydrous methanol and add the 6M HCl that AcCl prepares.

Step 7

The CH of amine hydrochlorate 10.07 ₂Cl ₂(200ml) solution Et ₃N (45.0ml, 315mmol) and Boc ₂(45.7g is 209mmol)-78 ℃ of processing for O.Reaction mixture at room temperature stirred spend the night, (2M, 200ml) dilution is extracted into CH with HCl ₂Cl ₂In.The organic layer that merges filters through dried over mgso, and vacuum concentration with chromatography purification (EtOAc/ hexane 1: 4), obtains hydroxy ester 10.08.

Step 8

Methyl ester 10.08 (3g, THF/H 10.5mmol) ₂O (1: 1) solution LiOHH ₂(645mg 15.75mmol) handles O, at room temperature stirs 2 hours.Reaction mixture aqueous hydrochloric acid (1M, 15ml) acidifying, vacuum concentration.The vacuum-drying resistates obtains 10.09 with quantitative output.

Step 9

Acid 10.09, CH that above-mentioned steps obtains ₂Cl ₂(50ml) and the solution NH of DMF (25ml) ₄Cl (2.94g, 55.5mmol), EDCI (3.15g, 16.5mmol), HOOBt (2.69g, 16.5mmol) and NMM (4.4g 44mmol) handles.Reaction mixture was at room temperature stirred 3 days.Solvent removed in vacuo, resistates is used dichloromethane extraction with the HCl aqueous solution (250ml) dilution.The organic layer that merges washs with saturated sodium bicarbonate aqueous solution, through dried over mgso, filters final vacuum and concentrates, and obtains 10.10, is directly used in next step.(perhaps, also can be by 10.06 (4.5g, 17.7mmol) and H ₂O ₂The aqueous solution (10ml), LiOHH ₂(820mg is 20.8mmol) at 50ml CH for O ₃In 0 ℃ of reaction 0.5 hour, directly obtain 10.10 among the OH).

Step 10

10.10 the solution that previous step is obtained is dissolved in 4N HCl/ two _ alkane, at room temperature stirs 2 hours.The vacuum concentration reaction mixture obtains solid intermediate 10.11, need not repurity and directly uses.

Step 11

Basically according to the method for describing in the above-mentioned steps 9,10, obtain required intermediate 10.12 with compound 10.09 and suitable reagent.

Preparation intermediate 11.01

Step 1

To 4-pentyne-1-alcohol 11.02 (4.15g; Aldrich) add Dess-Martin in the solution and cross iodine alkane (Periodinane) (30.25g; Aldrich), stir the gained mixture 45 minutes, add (tert-butoxycarbonyl methylene radical) triphenyl phosphorane (26.75g then; Aldrich).Stir gained black reaction thing and spend the night, with the EtOAc dilution, with sodium sulfite aqueous solution, saturated sodium bicarbonate aqueous solution, water, salt water washing, drying.Volatile matter is removed in decompression, and resistates obtains required compound 11.03 (3.92g) with silica gel column chromatography purifying (using the 1%EtOAc/ hexane as eluent).Also obtain some impure flow points, but temporarily put aside.

Step 2

Use the n-propyl alcohol (20ml of alkene 11.03 (1.9g); Aldrich) solution, benzyl carbamate (4.95g; Aldrich) water (79ml) solution of n-propyl alcohol (40ml) solution, NaOH (1.29g), t-butyl hypochlorate (3.7ml), (DHQ) ₂PHAL (0.423g; Aldrich) n-propyl alcohol (37.5ml) solution and potassium osmate dehydrate (0.1544g; Aldrich), according to Angew.Chem.Int.Ed.Engl (1998), 35, (23/24), the method of describing in the 2813-7 page or leaf obtains crude product, this crude product silica gel column chromatography purifying (EtOAc: hexane 1: 5), obtain required amino alcohol 11.04 (1.37g, 37%, white solid).

Step 3

In ester 11.04 (0.700g), add 4M HCl/ two _ alkane (20ml; Aldrich), gained mixture standing over night at room temperature.Volatile matter is removed in decompression, obtains acid 11.05 (0.621g, white solids).

Step 4

At room temperature, successively with bop reagent (3.65g; Sigma), triethylamine (3.45ml) joins in methylene dichloride (20ml) solution of carboxylic acid 11.05 (2.00g) and allylamine (0.616ml) stirring gained mixture overnight.Reaction mixture is distributed between the EtOAc and the 10%HCl aqueous solution.Isolate organic phase, with saturated sodium bicarbonate aqueous solution, water washing, through dried over mgso.Rough reaction product (is used EtOAc: hexane with the silica gel column chromatography purifying; 70: 30 as eluent), obtain required acid amides 11.01 (1.73g, yellow viscous oil shape thing).

Preparation intermediate 12.03 and 12.04

Step 1

Basically according to the method for describing among the intermediate 10.11 step 3-8, compound 12.01 is converted into desired substance 12.02.

Step 2

Basically according to the method for describing in intermediate 10.11 steps 9,10, compound 12.02 is converted into required intermediate 12.03.

Step 3

Basically according to the method for describing in intermediate 10.12 steps 11, compound 12.02 is converted into required intermediate 12.03.

Preparation intermediate 13.01 and 13.06

Step 1

At 0-5 ℃, in 2 hours to 1-nitrobutane 13.02 (16.5g, 0.16mol), the water of oxoethanoic acid (28.1g, 0.305mol) and drip in the stirred solution of MeOH (122ml) triethylamine (93ml, 0.667mol).Solution is risen to room temperature, and stirring is spent the night, and is concentrated into driedly, obtains oily matter.Oily matter is water-soluble, be acidified to pH=1 with 10%HCl, extract with EtOAc then.The organic solution salt water washing that merges, through dried over sodium sulfate, filtration is concentrated into driedly, obtains product 13.03 (28.1g, yield 99%).

Step 2

To compound 13.03 (240g, 1.35mol) and add 10%Pd/C (37g) in the stirred solution of acetate (1.25L).59psi hydrogenation gained solution 3 hours, spend the night in 60psi hydrogenation then.Evaporation of acetic acid with methylbenzene azeotropic 3 times, is ground with MeOH and ether then.Filtering solution with methylbenzene azeotropic 2 times, obtains pale solid 13.04 (131g, 0.891mol, 66%).

Step 3

To amino acid/11 3.04 (2.0g, 13.6mmol), add in 0 ℃ of stirred solution of two _ alkane (10ml) and water (5ml) 1N NaOH solution (4.3ml, 14.0mmol).Stirred gained solution 10 minutes, (0.110g 14.0mmol), stirred 15 minutes at 0 ℃ to add tert-Butyl dicarbonate then.Then solution is risen to room temperature, stirred 45 minutes, in refrigerator, preserve and spend the night, be concentrated into driedly, obtain crude product.In EtOAc of crude product (100ml) and ice solution, add KHSO ₄(3.36g) and water (32ml), stirred 4-6 minute.Isolate organic layer, water layer EtOAc extracting twice, the organic layer water of merging, salt water washing through dried over sodium sulfate, are filtered, and are concentrated into driedly, obtain product 13.05 (copal gum shape thing, 30g, yield 89%).

Step 4

Basically according to the method for describing in intermediate 10.12 steps 11, compound 13.05 is converted into required intermediate 13.01.

Step 5

Basically according to the method for describing in intermediate 10.12 steps 11, in coupled reaction, use cyclopropylamine (substituting allylamine), compound 13.05 is converted into required intermediate 13.06.

Preparation intermediate 14.01

Step 1

Basically according to the method for describing among the intermediate 13.01 step 1-3, compound 14.02 is converted into required product 14.03.

Step 2

Basically according to the method for describing in intermediate 10.12 steps 11, compound 14.03 is converted into required intermediate 14.01.

Preparation intermediate 15.01

Step 1

(9g, 58.5mmol) and in 0 ℃ of suspension of ether (25ml), slowly (about 15 minutes) add 4-iodo-1,1,1-trifluoro butane 15.02 (10g, ether 42.0mmol) (25ml) solution to silver nitrite by addition funnel.The gained mixture rises to room temperature 0 ℃ of vigorous stirring.After 50 hours, leach solid matter by Celite pad.Vacuum concentration gained diethyl ether solution obtains 15.03 colorless oil, need not repurity and directly uses.

Step 2

Basically according to the method for describing among the intermediate 13.01 step 1-3, compound 15.03 is converted into desired substance 15.04.

Step 3

Basically according to the method for describing in intermediate 10.12 steps 11, compound 15.04 is converted into required intermediate 15.01.

Preparation intermediate 16.01

Acid 16.02 (Winkler, D.; Burger, K., Synthesis, 1996,1419) (preparation intermediate 10.12) processing according to the method described above, obtain the intermediate 16.01 that needs.

Preparation intermediate 50.01

Step 1

In the MeOH of 50.02 (15g) (150ml) solution, add concentrated hydrochloric acid (3-4ml), mixture was refluxed 16 hours.Reaction mixture concentrates to room temperature.Resistates is dissolved in ether (250ml), with cold saturated sodium bicarbonate solution and salt water washing.Organic layer concentrates through dried over sodium sulfate, obtains methyl esters 50.03 (12.98g), is directly used in next step and need not repurity.

Step 2

Under nitrogen atmosphere, the methyl esters 50.03 that above-mentioned steps is obtained is dissolved in methylene dichloride (100ml), is cooled to-78 ℃.In 2 hours, drip DIBAL (the 1.0M dichloromethane solution, 200ml).Reaction mixture was risen to room temperature in 16 hours.Reaction mixture to 0 ℃ drips MeOH (5-8ml).Under agitation slowly add 10% sodium tartrate aqueous solutions of potassium (200ml).With methylene dichloride (100ml) dilution, isolate organic layer (following the part white precipitate).Organic layer, concentrates through dried over sodium sulfate with 1N HCl (250ml), salt solution (200ml) washing, obtains alcohol 50.04 (11.00g, clarification oily matter).

Step 3

The above alcohol that obtains 50.04 is dissolved in methylene dichloride (400ml), under nitrogen atmosphere, is cooled to 0 ℃.Add PCC (22.2g) in batches, reaction mixture was slowly risen to room temperature in 16 hours.Reaction mixture filters by Celite pad with ether (500ml) dilution.Concentrated filtrate is dissolved in ether (500ml) with resistates.Make it pass through silicagel pad, concentrated filtrate obtains aldehyde 50.05, is directly used in next step and need not repurity.

Step 4

Basically according to Chakraborty etc. (Tetrahedron, 1995,51 (33), method 9179-90) is converted into desired substance 50.01 with the above aldehyde that obtains 50.05.

Preparation intermediate 51.01

According to the method for describing in the document (T.K.Chakraborty etc., Tetrahedron, 1995,51 (33), 9179-90), obtain required intermediate 51.01 with aldehyde 51.02.

Preparation intermediate 60.01:

Step 1

In the 50ml anhydrous THF solution of phthalic imidine (1.01g), add triphenylphosphine (3eq) and N-tertbutyloxycarbonyl leucinol 60.02 (1eq).Mixture is cooled off in ice-water bath, drip diisopropyl azodiformate (2.5eq).Stirred the gained mixture 10 minutes at 0 ℃, rise to room temperature, stir about 2.5 hours is up to TLC (ethyl acetate/hexane; 3: 7) detect less than till any raw material.The concentrating under reduced pressure mixture.Resistates is suspended in the 80ml methylene dichloride.Leach solid.Concentrated filtrate adds hexane (30ml) to half volume.Leach solid.Concentrating under reduced pressure filtrate, resistates are used silica gel chromatography purifying (gradient: ethyl acetate/hexane; 1: 9 to 4: 6), obtain product 60.03.

Step 2

The amine 60.03 (1.4g) of N-Boc protection is dissolved in two _ alkane solution of 20ml 4M HCl.Stirred the mixture 2 hours.Vacuum is removed all volatile matters.Need not to be further purified, former state is used required product 60.04.

Step 3

0 ℃ of mixture of amine hydrochlorate 60.04 (1.14g), 20ml methylene dichloride and 20ml saturated sodium bicarbonate aqueous solution is handled with phosgene (10ml, 15% toluene solution), stirs 2 hours.Reaction mixture is with the dilution of 100ml methylene dichloride, with 30ml saturated sodium bicarbonate cold water solution washing.Organic layer filters through dried over mgso, uses the 10ml dilution with toluene again.Enriched mixture is preserved product 60.01 as the 0.2M toluene solution.

Preparation intermediate 61.01:

Step 1

According to the above-mentioned method that is used for intermediate 60.01, with 60.02 and 4,4-dimethyl-penten imide obtains required product 61.01.

Synthetic intermediate 62.01:

Step 1

At 0 ℃, to acid amides 62.02 (0.5g, add in THF solution 1eq) cyclopropyl bromination magnesium (4eq, 7.68mmol).After 15 minutes reactant is risen to room temperature, at room temperature reaction stirred is 5 hours, then by adding 1N HCl quencher.Reactant dilutes with EtOAc, uses the salt water washing.Organic layer with column chromatography purifying (10%EtOAc/ hexane), obtains 0.2g product 62.03 through dried over mgso.Yield 43.1%.

Step 2

Two _ alkane the solution that in the amine 62.03 (0.2g) of N-Boc protection, adds 4M HCl.At room temperature reaction stirred is 50 minutes, and TLC demonstration reaction is finished.Enriched mixture obtains the required product 62.04 of 0.162g to doing.

Step 3

At 0 ℃, to the CH of phosgene ₂Cl ₂Solution (2eq, 1.65mmol), add 62.04 in the mixture of sodium bicarbonate (5ml saturated aqueous solution).At room temperature stirred the mixture 2.5 hours.Isolate organic layer, through anhydrous sodium sulfate drying.Be concentrated into half volume with cooling bath.Be diluted to 10ml, obtain required isocyanic ester 62.01, be the 0.083M dichloromethane solution.

Synthetic intermediate 63.01:

Step 1

Under-78 ℃, nitrogen atmosphere, KHMDS (200ml 0.5M toluene solution) is added drop-wise to naphthenic acid methyl esters 63.02 (11.1g; 78mmol) and the stirred solution of anhydrous THF (200ml).After adding finishes, reactant is continued to maintain this temperature 0.5 hour.Add benzyl chloride methyl ether (18.6ml then; 134mmol).Allow reactant rise to ambient temperature overnight, add entry (100ml).The water law aftertreatment obtains resistates, and recycle silicon glue column chromatography purifying (use EtOAc: hexane is made eluent at 1: 10) obtains required impure intermediate ether (14.98g, colorless oil).

At room temperature, (black suspension of 4.1g and MeOH (80ml) is exposed to that (gas tank) spends the night under the nitrogen atmosphere with 10%Pd/C (0.5g), above-mentioned rough ether.By Celite pad filtering reaction thing, with solid methyl alcohol thorough washing.The filtrate that concentrating under reduced pressure merges, crude product are used silica gel column chromatography purifying (EtOAc: hexane 1: 5), obtain primary alconol (63.03; 0.62g, colorless oil).

Step 2

Under 0 ℃, nitrogen atmosphere, successively methylsulfonyl chloride (0.31ml) and triethylamine (0.75ml) are joined primary alconol (63.03, in stirred solution 0.62g).The gained mixture was stirred 0.5 hour under this temperature.Reaction mixture is extracted into EtOAc,,, concentrates through dried over mgso with 1M HCl, saturated sodium bicarbonate aqueous solution, water washing.(methanesulfonates 63.04 0.74g), is directly used in next step with it and need not repurity to obtain the yellow oily resistates.

Step 3

With

In ice bath the cooling and nitrogen atmosphere under, with dimethyl formamide (20ml; Anhydrous; Aldrich) join sodium hydride (0.56g; Aldrch) in, again tert.-butyl mercaptan is added in the suspension.Finish in case add, add methanesulfonates (63.04; Prepare according to above method with 2.00g alcohol; 63.03), the gained mixture at room temperature stirred spend the night.Reactant is distributed between EtOAc and water, isolate organic phase, through dried over mgso.Silica gel column chromatography (EtOAc: hexane 2: 98) handle, obtain methyl esters-sulfide (63.05; 1.75g).

EtOAc is added aqueous phase, add the 10%HCl aqueous solution up to water layer pH=1.Isolate organic layer, wash with water, drying, concentrating under reduced pressure obtains sulfide-formic acid (63.06; 0.747g, white solid).

Step 4

To sulfide (63.06; 2.287g) methyl alcohol (75ml) solution in add oxone solution (18.00g; Aldrich), at room temperature stirring the gained white suspension spends the night.Volatile matter is removed in decompression, and white solid is distributed between EtOAc and water.Isolate organic phase, drying concentrates, and obtains sulfone (63.07; 2.52g; Comprise partial solvent).

Step 5

Acid 63.07 (1.6lg) the 50ml toluene solution with DPPA (1eq, 1.33ml, d1.270) and triethylamine (1eq, 0.85ml, d0.726) processing.With mixture heating up to 100 ℃ 2 hours.Reaction mixture dilutes with saturated sodium bicarbonate aqueous solution, with methylene dichloride (2 * 100ml) extractions.The organic layer that merges saturated sodium bicarbonate aqueous solution and salt water washing.Organic layer filters through dried over mgso, and concentrating under reduced pressure is up to remaining about 20ml solvent.The solution of product 63.01 is adjusted to the isocyanic ester of 0.2M concentration with toluene.

Synthetic intermediate 64.01:

Step 1

In the 100ml MeOH of phthalic imidine 60.03 (g) solution, add hydrazine (0.9ml, 28.68mmol, 1.4eq), with mixture reflux (under nitrogen atmosphere) 6 hours.TLC confirms also to have part material, adds hydrazine (0.45ml) again, at room temperature continues to stir and spends the night.The adularescent precipitation generates.Leach solid, concentrated filtrate obtains white solid product 64.02 (4.48).

Step 2

(2.16g, 100ml dichloromethane solution 10mmol) is cooled to 0 ℃, with triethylamine (2eq, 2.8ml) processing with amine 64.02.The dropping methylsulfonyl chloride (1.2eq, 0.93ml).Stir heterogeneous mixture spend the night (at 0-25 ℃).Leach solid, filtrate is washed with saturated aqueous ammonium chloride (100ml) and salt solution (100ml).Organic layer filters the back and concentrates through dried over sodium sulfate.Resistates is dissolved in minimum dichloromethane/ethyl acetate (about 10ml), leaches insoluble white solid.Filtrate is used the silica gel column chromatography purifying, obtains product 64.03 (2.7g, thickness semisolid).

Step 3

(2.2g, 50ml anhydrous DMF solution 7.5mmol) is cooled to 0 ℃, with cesium carbonate (3eq, 7.34g) processing with sulphonamide 64.03.(5eq 2.34ml), stirred the mixture 45 minutes to drip methyl iodide.Remove cooling bath, restir mixture 4 hours.By adding saturated aqueous ammonium chloride (100ml) quencher reactant, with ethyl acetate (2 * 100ml) extractions.The organic layer water (200ml) that merges, salt solution (200ml) washing are through dried over sodium sulfate.Organic layer is filtered the back to be concentrated.Resistates is handled with silica gel chromatography, obtains product 64.04 (2.16g).

Step 4

At room temperature, (2.1g 6.82mmol) is dissolved in 20ml4M HCl/ two _ alkane to the amine 64.04 that N-Boc is protected.Stirred reaction mixture 1 hour, all volatile matters are removed in decompression then, obtain product 64.05 with quantitative output.

Step 5

0 ℃ of mixture of amine hydrochlorate 64.05, methylene dichloride and saturated sodium bicarbonate aqueous solution is handled with phosgene (15% toluene solution), stirred 2 hours.Reaction mixture dilutes with methylene dichloride, with cold saturated sodium bicarbonate aqueous solution washing.Organic layer filters through dried over mgso, uses dilution with toluene again.Enriched mixture is regulated product 64.01, preserves as the 0.2M toluene solution.

Synthetic intermediate 65.01:

Step 1

Isocyanic ester 65.01 according to the method preparation that is used for isocyanic ester 64.01, is substituted methylsulfonyl chloride with 2-thiophene SULPHURYL CHLORIDE in the sulphonamide synthesis step.

Preparation embodiment's is synthetic

Synthetic embodiment 101

Step 1

To proline derivative 1.01 (3.66mmol, preparation according to the method described above), in 0 ℃ of stirred solution of methylene dichloride (20ml) and DMF (15ml), add L-tertbutyloxycarbonyl-Terleu (930mg, 4.03mmol), DIPEA (2.02ml, 10.98mmol) and HATU (1.8g, 4.76mmol).After keeping 15 minutes under this temperature, reaction flask is placed on spend the night in the refrigerator (20 ℃) (16 hours).Reaction mixture,, filters the back and concentrates through dried over sodium sulfate with saturated sodium bicarbonate solution (80ml), 10% aqueous citric acid solution (80ml), salt solution (80ml) washing with methylene dichloride (80ml) dilution.Crude product obtains the required product 101a of 1.77g with silica gel chromatography purifying (25/75 to 50/50 EtOAc/ hexane).LC-MS：518.1(M+H) ⁺。

Step 2

To methyl esters 101a (1.21g, 2.34mmol), add the 1M LiOH aqueous solution (5ml) in the solution of THF (10ml) and MeOH (5ml).Reaction mixture was at room temperature stirred 4 hours.Concentrate then, water (50ml) dilution is after separating out white solid matter, with solid citric acid acidifying (pH is about 3).Leach solid, wash with water, vacuum-drying obtains 970mg101b.LC-MS：504.1(M+H) ⁺。

Step 3

Basically according to above method (step 1, preparation 101a), (503mg, 1mmol) (334mg, 1.5mmol) coupling obtains 101c, directly uses to need not purifying with intermediate 13.06 with sour 101b.MS：672.37(M+H) ⁺。

Step 4

Adding Dess-Martin in the solution of above oxy-compound 101c and methylene dichloride (15ml) crosses iodine alkane (periodinane) (848mg 2mmol), at room temperature stirred reaction mixture 5 hours.At this moment, reaction mixture,, filters the back and concentrates through dried over sodium sulfate with 1: 1 mixture (2 times, each 25ml), salt solution (50ml) washing of 10% sodium thiosulfate solution and saturated sodium bicarbonate solution with methylene dichloride (30ml) dilution.Crude product obtains the required product 101d of 410mg with silica gel chromatography purifying (15/85 to 50/50 acetone/hexane).LC-MS：670.2(M+H) ⁺。

Step 5

According to the method that is used for intermediate 1.01 steps 3 (reaction times=2 hour), slough the protection of N-boc functional group among the 101d, obtain desired substance 101e.LC-MS：570.1(M+H) ⁺。

Step 6

To amine salt 101e (60mg, 0.1mmol) and in 0 ℃ of solution of methylene dichloride (2ml), add successively DIPEA (0.06ml, 0.3mmol), isocyanic ester intermediate 65.01 (the 0.25M toluene solution, 0.8ml, 0.2mmol).After keeping 15 minutes under this temperature, reaction flask is placed on spend the night in the refrigerator (20 ℃) (16 hours).Reaction mixture,, filters the back and concentrates through dried over sodium sulfate with saturated ammonium chloride solution (20ml), salt solution (20ml) washing with methylene dichloride (20ml) dilution.Crude product obtains required compound 101 (53mg) with silica gel chromatography purifying (15/85 to 50/50 acetone/hexane); LC-MS:872.2 (M+H) ⁺

Synthetic embodiment 102

Step 1

According to the method for embodiment 101 steps 6, with 101e and intermediate 63.01 preparation required compounds 102.102 LC-MS:829.2 (M+H) ⁺

Synthetic embodiment 103

Step 1

According to the method for embodiment 101 steps 6, with 101e and intermediate 64.01 preparation required compounds 103.103 LC-MS:804.2 (M+H) ⁺

According to above-mentioned similar approach, other compound in the preparation table 2.In addition, the compound in the table 1 also can prepare similarly.

The present invention relates to new HCV proteinase inhibitor.This effectiveness can be proved by the ability that they suppress HCV NS3/NS4a serine protease.The general method that is used for described proof illustrates by following external test method.

The test of HCV protease inhibiting activity:

Spectrophotometry: according to R.Zhang etc. (Analytical Biochemistry, 270(1999) 268-275, its disclosure is attached to herein by reference) method described, The compounds of this invention is carried out the spectrophotometry of HCV serine protease.Mensuration based on proteolysis colour developing ester substrate is fit to continuous monitoring HCV NS3 protease activity.Substrate is derived from the P side of NS5A-NS5B catenation sequence (Ac-DTEDVVX (Nva), wherein X=A or P), and its C end carboxyl is by one of 4 different color development alcohol (3-nitrophenol, 4-nitrophenol, 7-hydroxy-4-methyl-tonka bean camphor or 4-phenylazo phenol) esterification.Introduce these new spectrophotometrys below and use synthesizing, characterizing and the application in high flux screening of ester substrate, and introduce the kinetics evaluation of HCV NS3 proteinase inhibitor in detail.

Materials and methods:

Material: measure with the chemical reagent in the relevant damping fluid available from Sigma ChemicalCompany (St.Louis, Missouri).The synthetic reagent of using of peptide is available from Aldrich Chemicals, Novabiochem (San Diego, California), Applied Biosystems (Foster City, California) and Perseptive Biosystems (Framingham, Massachusetts).The artificial synthesis peptide perhaps uses automatic ABI 431A type synthesizer (Applied Biosystems) to synthesize peptide.(Norwalk, Connecticut), 96 hole UV plates are available from Corning (Corning, New York) available from Perkin Elmer for LAMBDA 12 type UV/VIS spectrometers.Preheating table (prewarming block) available from USA Scientific (Ocala, Florida), 96 orifice plate vortices available from Labline Instruments (Melrose Park, Illinois).Spectramax Plus microtiter plate readout instrument (having monochrometer) available from Molecular Devices (Sunnyvale, California).

The enzyme preparation: use previously disclosed method (D.L.Sali etc., Biochemistry, 37(1998) preparation recombinant HCV NS3/NS4A heterodimer proteolytic enzyme (bacterial strain la) 3392-3401).Use in advance by the quantitative recombinant HCV proteolytic enzyme of amino acid analysis standard substance, by Biorad dye method mensuration protein concn.Before beginning to measure, adopt Biorad Bio-Spin P-6 prepacked column, enzyme storage buffer (50mM sodium phosphate pH8.0,300mM sodium-chlor, 10% glycerine, 0.05% lauryl maltoside and 10mM DTT) is exchanged for measures damping fluid (25mMMOPS pH6.5,300mM sodium-chlor, 10% glycerine, 0.05% lauryl maltoside, 5 μ M EDTA and 5 μ M DTT).

Synthetic and the purifying of substrate: according to the synthetic substrate of report such as R.Zhang (the same), during synthetic beginning with standard method (K.Barlos etc., Int.J.Pept.Protein Res., 37(1991), 513-520), Fmoc-Nva-OH is anchored on the 2-chlorine trityl chloride resin.Carry out subsequently or manually or adopt automatic ABI 431 type peptide synthesizers, utilize Fmoc chemistry secretory piece.The peptide fragment of N-acetylize and protection fully was through methylene dichloride (DCM) solution-treated of 10% acetate (HOAc) and 10% trifluoroethanol (TFE) 30 minutes, and perhaps the DCM solution-treated through 2% trifluoroacetic acid (TFA) disconnected with resin after 10 minutes.Filtrate and DCM washings (perhaps repeating to extract) that azeotropic vaporization merges with aqueous sodium carbonate, the acid of using when removing cracking.DCM evaporates after dried over sodium sulfate.

Described ester substrate standard acid-pure couling process (K.Holmber etc., Acta Chem.Scand, B33(1979) 410-412) assemble.Peptide fragment is dissolved in anhydrous pyridine (30-60mg/ml), to wherein adding 10 molar equivalent chromophoric grouies and catalytic amount (0.1eq.) tosic acid (pTSA).(DCC 3eq.) starts coupled reaction to add dicyclohexylcarbodiimide.Form with HPLC monitoring product, confirm that at room temperature reacting 12-72 hour afterreaction finishes.Pyridine solvent is removed by vacuum-evaporation, further removes with methylbenzene azeotropic.The DCM solution deprotection of peptide ester usefulness 95%TFA 2 hours extracts 3 times to remove excessive chromophoric group with anhydrous diethyl ether.The deprotection substrate by the reversed-phase HPLC purifying, is used 30%-60% acetonitrile gradient wash-out (with 6 column volumes) on C3 or C8 post.Total recovery behind the HPLC purifying is about 20-30%.The electro-spray ionization mass spectrum confirms molecular weight.Substrate is preserved so that dry powder form is moistureproof.

The spectrum of substrate and product: measure the spectrum that damping fluid obtains substrate and corresponding chromophoric group product with pH6.5.Utilize a plurality of extent of dilution, the 1cm cuvette, optimum non-peak wavelength (3Np and HMC are 340nm, and PAP is 370nm, and 4-Np is 400nm) is measured optical extinction coefficient.Optimum non-peak wavelength is defined as: the wavelength that produces maximum absorbance mark poor ((product OD-substrate OD)/substrate OD) between substrate and the product.

Protease assay: in 96 hole microtiter plates,, carry out the HCV protease assay in 30 ℃ with 200 μ l reaction mixtures.To the NS3/NS4A heterodimer, optimize and measure buffer condition (25mM MOPS pH6.5,300mM NaCl, 10% glycerine, 0.05% lauryl maltoside, 5 μ M EDTA and 5 μ M DTT) (D.L.Sali etc., ibid).Usually, 150 μ l mixtures of buffer reagent, substrate and inhibitor are added in each hole (the DMSO final concentration≤4%v/v), about 3 minutes of 30 ℃ of pre-incubations.Start reaction (final volume is 200 μ l) with the pre-hot solution of proteolytic enzyme (12nM, 30 ℃) of 50 μ l in measuring damping fluid then.With the Spectromax Plus microtiter plate readout instrument that is equipped with monochromator (adopting edge filter template readout instrument can obtain acceptable result), (3Np and HMC are 340nm at suitable wavelength, PAP is 370nm, 4-Np is 400nm), the absorbancy of each plate of monitoring changes in minute (60 minutes) scope.With respect to no enzyme blank as no enzymic hydrolysis contrast, the proteolysis of the ester bond between suitable wavelength monitoring Nva and chromophoric group.(about 6-200 μ M) estimates the substrate kinetics parameter in 30 times of concentration of substrate scopes.Obtain initial velocity with linear regression, (Mac Curve Fit 1.1 K.Raner), is fitted to the Michaelis-Menten equation with experimental data, draws kinetic constant to adopt nonlinear regression analysis.Calculate enzyme turnover ratio (k _Cat), suppose that enzyme has complete activity.

The evaluation of inhibitor and inactivator: to striving property inhibitor Ac-D-(D-Gla)-L-I-(Cha)-C-OH (27), Ac-DTEDVVA (Nva)-OH and Ac-DTEDVVP (Nva)-OH unexpectedly, according to being used for the dynamic (dynamical) rearrangement of striving property inhibition Michaelis-Menten equation unexpectedly: v _o/ v _i=1+[I] _o/ (K _i(1+[S] _o/ K _m)), v wherein _oInitial velocity when not suppressing, v _iBe any known inhibitor concentration ([I] _o) initial velocity when inhibitor exists, [S] _oBe employed concentration of substrate; Use v _o/ v _iTo inhibitor concentration ([I] _o) mapping, through the enzyme of measuring fixed concentration and the inhibition constant (K of substrate _i).With the data that linear regression fit obtains, use the slope 1/ (K that is drawn _i(1+[S] _o/ K _m), calculating K _iValue.Provided the Ki of part of compounds of the present invention in the table 2 ^*Value (nM of unit).

Table 2

Although illustrated the present invention in conjunction with above-mentioned specific embodiments, for those of ordinary skills, many replacements of the present invention, modification and other variation scheme are conspicuous.All such replacements, modification and variation scheme all fall within the spirit and scope of the invention.

Claims

1. the pharmacy acceptable salt of the enantiomer of a compound or described compound, steric isomer, rotational isomer, tautomer and racemic modification or described compound or solvate, described compound has the structure of following general formula I:

Formula I

Wherein:

R ¹Be H, OR ⁸, NR ⁹R ¹⁰Or CHR ⁹R ¹⁰, R wherein ⁸, R ⁹And R ¹⁰Can be identical or different, and independently be selected from separately H, alkyl-, thiazolinyl-, alkynyl-, aryl-, assorted alkyl-, heteroaryl-, cycloalkyl-, heterocyclic radical-, arylalkyl-and heteroarylalkyl, perhaps NR ⁹R ¹⁰In R ⁹And R ¹⁰Interconnect, make NR ⁹R ¹⁰Constitute 4-8 unit heterocyclic radical, same, CHR ⁹R ¹⁰In R ⁹And R ¹⁰Also interconnect, make CHR ⁹R ¹⁰Constitute 4-8 unit cycloalkyl;

Y is selected from following part:

2. the compound of claim 1, wherein said compound has the following formula structure:

Wherein definition in various piece such as the claim 1.

3. the compound of claim 1, wherein R ¹Be NR ⁹R ¹⁰, R ⁹Be H, R ¹⁰Be H or R ¹⁴, R wherein ¹⁴Be alkyl, aryl, assorted alkyl, heteroaryl, cycloalkyl, alkyl-aryl, alkyl-heteroaryl, aryl-alkyl, thiazolinyl, alkynyl or heteroaryl-alkyl.

4. the compound of claim 3, wherein R ¹⁴Be selected from following part:

5. the compound of claim 1, wherein R ²Be selected from following part:

6. the compound of claim 1, wherein R ³Be selected from following part:

R wherein ³¹Be OH or O-alkyl;

R ³²Be H, C (O) CH ₃, C (O) O ^tBu or C (O) N (H) ^tBu.

7. the compound of claim 6, wherein R ³Be selected from following part:

8. the compound of claim 1, wherein G is NH.

9. the compound of claim 8, wherein Y is selected from following part:

R wherein ¹⁵, R ¹⁶, R ¹⁷, R ¹⁸, R ¹⁹, R ²⁰, R ²¹, R ²², R ²³, R ²⁴And R ²⁵Independently be selected from H, alkyl, assorted alkyl, thiazolinyl, assorted thiazolinyl, alkynyl, assorted alkynyl, cycloalkyl, heterocyclic radical, aryl, arylalkyl, heteroaryl and heteroarylalkyl, perhaps (i) R separately ¹⁷And R ¹⁸Connect and compose 3-8 unit's cycloalkyl or heterocyclic radical independently of one another; (ii) R ¹⁵And R ¹⁹Connect and compose 4-8 unit heterocyclic radical independently of one another; (iii) R ¹⁵And R ¹⁶Connect and compose 4-8 unit heterocyclic radical independently of one another; (iv) R ¹⁵And R ²⁰Connect and compose 4-8 unit heterocyclic radical independently of one another;

10. the compound of claim 9, wherein following part:

Be selected from:

With

Y wherein ³²Be selected from following groups:

With

11. the compound of claim 9, wherein Y is selected from:

Or

12. the compound of claim 1, wherein Z is selected from following part:

13. the compound of claim 12, wherein Z is selected from following part:

14. the compound of claim 13, wherein Z is selected from following part:

15. the compound of claim 1, wherein R ¹Be NH ₂Or NHR ¹⁴, R wherein ¹⁴Be selected from following part:

With

R ²Be selected from following part:

R ³Be selected from following part:

Z is selected from:

Y is selected from:

16. a pharmaceutical composition, said composition comprises the compound as at least a claim 1 of activeconstituents.

17. the pharmaceutical composition of claim 16, said composition are used for the treatment of hepatitis C virus (HCV) relative disease.

18. the pharmaceutical composition of claim 17, said composition also comprise at least a pharmaceutically acceptable carrier.

19. the pharmaceutical composition of claim 18, said composition also comprise at least a antiviral drug.

20. the pharmaceutical composition of claim 19, said composition also comprise at least a Interferon, rabbit.

21. the pharmaceutical composition of claim 20, wherein said at least a antiviral drug is a ribavirin, and described at least a Interferon, rabbit is interferon-alpha or the Interferon, rabbit that adds polyoxyethylene glycol.

22. the compound of at least a claim 1 of treatment significant quantity is used for the treatment of purposes in the medicine of hepatitis C virus (" HCV ") relative disease in preparation.

23. the purposes of claim 22, wherein said medicine is fit to oral or subcutaneous administration.

24. compound or enantiomer, steric isomer, rotational isomer, tautomer and the racemic modification of described compound or the pharmacy acceptable salt or the solvate of described compound with HCV protease inhibiting activity, described compound is selected from the compound with following array structure:

25. compound or enantiomer, steric isomer, rotational isomer, tautomer and the racemic modification of described compound or the pharmacy acceptable salt or the solvate of described compound with HCV protease inhibiting activity, described compound is selected from the compound with following array structure:

26. a pharmaceutical composition for the treatment of the HCV relative disease, described composition comprise the compound and the pharmaceutically acceptable carrier of one or more claims 24 for the treatment of significant quantity.

27. the pharmaceutical composition of claim 26, said composition also comprise at least a antiviral drug.

28. the pharmaceutical composition of claim 27, said composition also comprise at least a Interferon, rabbit or polyoxyethylene glycol (PEG)-interferon alpha conjugate.

29. the pharmaceutical composition of claim 28, wherein said at least a antiviral drug is a ribavirin, and described at least a Interferon, rabbit is interferon-alpha or the Interferon, rabbit that adds polyoxyethylene glycol.

30. the compound of at least a claim 24 of treatment significant quantity is used for the treatment of purposes in the medicine of hepatitis C virus relative disease in preparation.

31. the compound of at least a claim 24 of treatment significant quantity is used for regulating the purposes of the medicine of hepatitis C virus (HCV) protease activity in preparation.

32. the compound of at least a claim 24 of treatment significant quantity is used for the treatment of, prevents in preparation or improves purposes in the medicine of one or more symptoms of hepatitis C virus.

33. the purposes of claim 31, wherein said HCV proteolytic enzyme is NS3/NS4a proteolytic enzyme.

34. the purposes of claim 33, wherein said one or more compounds suppress HCVNS3/NS4a proteolytic enzyme.

35. the compound of at least a claim 24 of treatment significant quantity is used for regulating the purposes of the medicine of hepatitis C virus (HCV) polypeptide processing in preparation.

36. the pharmaceutical composition of treatment significant quantity is used for the treatment of purposes in the medicine of HCV relative disease in preparation, described pharmaceutical composition comprises at least a compound or enantiomer, steric isomer, rotational isomer, tautomer and the racemic modification of described compound or the pharmacy acceptable salt or the solvate of described compound for the treatment of significant quantity, and described compound is selected from following compounds:

37. the compound of the claim 1 of purified form.